{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T13:14:15Z","timestamp":1793193255580,"version":"4.5.0"},"reference-count":42,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2020,2,11]],"date-time":"2020-02-11T00:00:00Z","timestamp":1581379200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000776","name":"Orthopaedic Research UK","doi-asserted-by":"publisher","award":["ref 495 (15082012)"],"award-info":[{"award-number":["ref 495 (15082012)"]}],"id":[{"id":"10.13039\/501100000776","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>To characterise the dynamic of events during the early phases of fracture repair in humans, we investigated molecular events using gene expression profiling of bone fragments from the fracture site at different time points after trauma and immune\/stromal cells recruitment at the fracture site using flow cytometry. Bone and inflammatory markers were expressed at low levels at homeostasis, while transcripts for bone constituent proteins were consistently detected at higher levels. Early after fracture (range 2\u20134 days), increased expression of CXCL12, suggested recruitment of immune cells associated with a change in the balance of degradation enzymes and their inhibitors. At intermediate time after fracture (4\u20138 days), we observed high expression of inflammatory cytokines (IL1-beta, IL6), CCL2, the T-cell activation marker CD69. Late after fracture (8\u201314 days), high expression of factors co-operating towards the regulation of bone turnover was detected. We identified potential soluble factors and explored circulating levels in patients for whom a union\/non-union (U\/NU) outcome was known. This showed a clear difference for PlGF (p = 0.003) at day 1. These findings can inform future studies further investigating the cascade of molecular events following fractures and for the prediction of fracture non-union.<\/jats:p>","DOI":"10.3390\/jcm9020492","type":"journal-article","created":{"date-parts":[[2020,2,11]],"date-time":"2020-02-11T11:45:30Z","timestamp":1581421530000},"page":"492","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Dynamics of Early Signalling Events during Fracture Healing and Potential Serum Biomarkers of Fracture Non-Union in Humans"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3321-9087","authenticated-orcid":false,"given":"Agata N.","family":"Burska","sequence":"first","affiliation":[{"name":"Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS2 9JT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter V.","family":"Giannoudis","sequence":"additional","affiliation":[{"name":"Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS2 9JT, UK"},{"name":"Leeds Biomedical Research Centre, Leeds Teaching Hospitals NHS Trust (LTHT), Leeds LS9 7TF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5055-7630","authenticated-orcid":false,"given":"Boon Hiang","family":"Tan","sequence":"additional","affiliation":[{"name":"Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS2 9JT, UK"},{"name":"Leeds Biomedical Research Centre, Leeds Teaching Hospitals NHS Trust (LTHT), Leeds LS9 7TF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6404-6620","authenticated-orcid":false,"given":"Dragos","family":"Ilas","sequence":"additional","affiliation":[{"name":"Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS2 9JT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9365-2283","authenticated-orcid":false,"given":"Elena","family":"Jones","sequence":"additional","affiliation":[{"name":"Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS2 9JT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3969-7701","authenticated-orcid":false,"given":"Frederique","family":"Ponchel","sequence":"additional","affiliation":[{"name":"Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS2 9JT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,2,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S3","DOI":"10.1016\/S0020-1383(08)70003-2","article-title":"Fracture healing: The diamond concept","volume":"38","author":"Giannoudis","year":"2007","journal-title":"Injury"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1392","DOI":"10.1016\/j.injury.2005.07.019","article-title":"Current concepts of molecular aspects of bone healing","volume":"36","author":"Dimitriou","year":"2005","journal-title":"Injury"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"S32","DOI":"10.1016\/j.injury.2014.04.006","article-title":"Biomechanical methods for the assessment of fracture repair","volume":"45","author":"Augat","year":"2014","journal-title":"Injury"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ekegren, C., Edwards, E., de Steiger, R., and Gabbe, B. (2018). Incidence, costs and predictors of non-union, delayed union and mal-union following long bone fracture. Int. J. Environ. Res. Public Health, 15.","DOI":"10.3390\/ijerph15122845"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.bone.2014.07.033","article-title":"Bone fracture healing: Cell therapy in delayed unions and nonunions","volume":"70","author":"Rosset","year":"2015","journal-title":"Bone"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1111\/jcmm.12532","article-title":"Biological and molecular profile of fracture non-union tissue: Current insights","volume":"19","author":"Panteli","year":"2015","journal-title":"J. Cell. Mol. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.afos.2015.09.004","article-title":"Communication between the skeletal and immune systems","volume":"1","author":"Lee","year":"2015","journal-title":"Osteoporos. Sarcopenia"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1002\/jor.20901","article-title":"Cellular composition of the initial fracture hematoma compared to a muscle hematoma: A study in sheep","volume":"27","author":"Schell","year":"2009","journal-title":"J. Orthop. Res."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kovtun, A., Messerer, D.A., Scharffetter-Kochanek, K., Huber-Lang, M., and Ignatius, A. (2018). Neutrophils in tissue trauma of the skin, bone, and lung: Two sides of the same coin. J. Immunol. Res., 2018.","DOI":"10.1155\/2018\/8173983"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1007\/s11914-018-0423-2","article-title":"The role of the immune cells in fracture healing","volume":"16","author":"Baht","year":"2018","journal-title":"Curr. Osteoporos. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/j.injury.2011.03.031","article-title":"The biology of fracture healing","volume":"42","author":"Marsell","year":"2011","journal-title":"Injury"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Edderkaoui, B. (2017). Potential role of chemokines in fracture repair. Front. Endocrinol., 8.","DOI":"10.3389\/fendo.2017.00039"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1038\/nrrheum.2012.185","article-title":"Correction: Screening: Cytof\u2014The next generation of cell detection","volume":"8","author":"Cheung","year":"2012","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1007\/s00109-004-0612-6","article-title":"Mechanistic insight into osteoclast differentiation in osteoimmunology","volume":"83","author":"Takayanagi","year":"2005","journal-title":"J. Mol. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4104","DOI":"10.1002\/art.23138","article-title":"Treg cells suppress osteoclast formation: A new link between the immune system and bone","volume":"56","author":"Zaiss","year":"2007","journal-title":"Arthritis Rheum."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1089\/ten.teb.2009.0687","article-title":"The early fracture hematoma and its potential role in fracture healing","volume":"16","author":"Kolar","year":"2010","journal-title":"Tissue Eng. Part B Rev."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1694","DOI":"10.1002\/art.24520","article-title":"A role for \u03b3\/\u03b4 t cells in a mouse model of fracture healing","volume":"60","author":"Colburn","year":"2009","journal-title":"Arthritis Rheum."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1038\/nrrheum.2012.1","article-title":"Fracture healing under healthy and inflammatory conditions","volume":"8","author":"Claes","year":"2012","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1097\/BOT.0b013e3182251421","article-title":"Preclinical animal models in trauma research","volume":"25","author":"Harvey","year":"2011","journal-title":"J. Orthop. Trauma"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1159\/000358663","article-title":"Inflammatory microenvironment changes the secretory profile of mesenchymal stem cells to recruit mesenchymal stem cells","volume":"33","author":"Xing","year":"2014","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2632","DOI":"10.1002\/art.34434","article-title":"Transcriptional profile of native cd271+ multipotential stromal cells: Evidence for multiple fates, with prominent osteogenic and wnt pathway signaling activity","volume":"64","author":"Churchman","year":"2012","journal-title":"Arthritis Rheum."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1097\/01.sla.0000171866.45848.68","article-title":"Osteonectin influences growth and invasion of pancreatic cancer cells","volume":"242","author":"Guweidhi","year":"2005","journal-title":"Ann. Surg."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4473","DOI":"10.1073\/pnas.87.12.4473","article-title":"Osteopontin\u2014A possible anchor of osteoclasts to bone","volume":"87","author":"Reinholt","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1731","DOI":"10.1126\/science.1062960","article-title":"The influence of the proinflammatory cytokine, osteopontin, on autoimmune demyelinating disease","volume":"294","author":"Chabas","year":"2001","journal-title":"Science"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1159\/000072956","article-title":"The roles of soluble osteopontin using osteopontin-transgenic mice In Vivo: Proliferation of cd4+ t lymphocytes and the enhancement of cell-mediated immune responses","volume":"71","author":"Higuchi","year":"2004","journal-title":"Pathobiol. J. Immunopathol. Mol. Cell. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1007\/s00223-003-0117-2","article-title":"Quantification, localization, and expression of igf-i and tgf-\u03b21 during growth factor-stimulated fracture healing","volume":"74","author":"Wildemann","year":"2004","journal-title":"Calcif. Tissue Int."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1016\/j.bone.2013.08.009","article-title":"Induced periosteum a complex cellular scaffold for the treatment of large bone defects","volume":"57","author":"Cuthbert","year":"2013","journal-title":"Bone"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3858\/emm.2012.44.1.025","article-title":"The discovery of placenta growth factor and its biological activity","volume":"44","year":"2012","journal-title":"Exp. Mol. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1230","DOI":"10.1172\/JCI26772","article-title":"Placental growth factor mediates mesenchymal cell development, cartilage turnover, and bone remodeling during fracture repair","volume":"116","author":"Maes","year":"2006","journal-title":"J. Clin. Investig."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2420","DOI":"10.1002\/stem.1482","article-title":"Orchestrating osteogenic differentiation of mesenchymal stem cells\u2014Identification of placental growth factor as a mechanosensitive gene with a pro-osteogenic role","volume":"31","author":"McCoy","year":"2013","journal-title":"Stem Cells"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/j.bone.2013.02.010","article-title":"Effects of pharmacological inhibition of cathepsin k on fracture repair in mice","volume":"55","author":"Gentile","year":"2013","journal-title":"Bone"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1016\/j.bone.2005.06.015","article-title":"Matrix metalloproteinases and failed fracture healing","volume":"37","author":"Henle","year":"2005","journal-title":"Bone"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1242\/dmm.006304","article-title":"Impaired remodeling phase of fracture repair in the absence of matrix metalloproteinase-2","volume":"4","author":"Lieu","year":"2011","journal-title":"Dis. Model. Mech."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1089\/ten.teb.2014.0677","article-title":"Boon and bane of inflammation in bone tissue regeneration and its link with angiogenesis","volume":"21","author":"Kwee","year":"2015","journal-title":"J. Tissue Eng. Part B Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1007\/s12026-016-8868-9","article-title":"Immunological characterization of the early human fracture hematoma","volume":"64","author":"Hoff","year":"2016","journal-title":"Immunol. Res."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Hoff, P., Gaber, T., Strehl, C., Jakstadt, M., Hoff, H., Schmidt-Bleek, K., Lang, A., Rohner, E., Huscher, D., and Matziolis, G. (2017). A pronounced inflammatory activity characterizes the early fracture healing phase in immunologically restricted patients. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18030583"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1007\/s12026-011-8235-9","article-title":"Immunologically restricted patients exhibit a pronounced inflammation and inadequate response to hypoxia in fracture hematomas","volume":"51","author":"Hoff","year":"2011","journal-title":"Immunol. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1002\/jcb.10435","article-title":"Fracture healing as a post-natal developmental process: Molecular, spatial, and temporal aspects of its regulation","volume":"88","author":"Gerstenfeld","year":"2003","journal-title":"J. Cell. Biochem."},{"key":"ref_39","first-page":"559","article-title":"Role of angiogenesis on bone formation","volume":"27","author":"Lozano","year":"2012","journal-title":"Histol. Histopathol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1016\/j.biopha.2018.06.078","article-title":"Angiopoietin 2 promotes angiogenesis in tissue-engineered bone and improves repair of bone defects by inducing autophagy","volume":"105","author":"Yin","year":"2018","journal-title":"Biomed. Pharmacother."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1016\/j.injury.2011.03.055","article-title":"Elevated transforming growth factor-beta 1 (tgf-\u03b21) levels in human fracture healing","volume":"42","author":"Sarahrudi","year":"2011","journal-title":"Injury"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1002\/jor.20906","article-title":"Vegf serum concentrations in patients with long bone fractures: A comparison between impaired and normal fracture healing","volume":"27","author":"Sarahrudi","year":"2009","journal-title":"J. Orthop. Res."}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/9\/2\/492\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T08:56:55Z","timestamp":1760173015000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/9\/2\/492"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,2,11]]},"references-count":42,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2020,2]]}},"alternative-id":["jcm9020492"],"URL":"https:\/\/doi.org\/10.3390\/jcm9020492","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,2,11]]}}}